外阴阴道假丝酵母菌病患者阴道乳杆菌的功能变化

Function of Lactobacillus in the Vagina of Vulvovaginal Candidiasis

目的分离、纯化、鉴定外阴阴道假丝酵母菌病(VVC)患者阴道乳杆菌菌种;绘制VVC患者阴道内分离的卷曲乳杆菌、加氏乳杆菌、詹氏乳杆菌、发酵乳杆菌的生长曲线,检测其产酸能力、产H_2O_2能力,分析VVC患者阴道乳杆菌的功能变化及临床意义,为VVC的病因及治疗提供理论基础。方法体外培养VVC患者阴道分泌物中的乳杆菌,分离、纯化,并用分子生物学技术鉴定;测定24h菌液浊度法绘制VVC患者及健康女性阴道内分离的4种乳杆菌的生长曲线;检测24h菌液p H值对比VVC患者与健康女性阴道内分离的卷曲乳杆菌、加氏乳杆菌、詹氏乳杆菌、发酵乳杆菌的产酸能力;绘制H_2O_2浓度标准曲线,定量检测摇菌3h菌液的产H_2O_2的量,对比VVC患者与健康女性阴道内分离的卷曲乳杆菌、加氏乳杆菌、詹氏乳杆菌、发酵乳杆菌的产H_2O_2能力。结果共分离培养了34株VVC患者阴道乳杆菌,经鉴定,分属于乳杆菌的4个种。VVC患者与健康妇女相比,阴道内卷曲乳杆菌、加氏乳杆菌、詹氏乳杆菌、发酵乳杆菌的增殖速度差异无统计学意义(P<0.05)。健康女性加氏乳杆菌与卷曲乳杆菌的产酸能力比VVC患者强(P<0.05);詹氏乳杆菌的产酸能力两组差异无统计学意义(P>0.05);而VVC患者发酵乳杆菌的产酸能力比健康女性强(P<0.05)。VVC患者阴道内分离的卷曲乳杆菌、加氏乳杆菌、詹氏乳杆菌、发酵乳杆菌的产H_2O_2量均明显少于健康妇女(P<0.05)。结论本研究为进一步构建中国女性阴道来源的乳杆菌菌种库、进行阴道来源的乳杆菌的相关研究提供菌种。VVC状态下,念珠菌的增多,阴道微环境的改变,并未影响阴道内定植的乳杆菌的生长速度。阴道内乳杆菌的产酸功能并不是抑制VVC的关键。阴道内乳杆菌产H_2O_2能力可能为抑制VVC的关键。

Objective To isolate, purificate, identificate lactobacillus strains in the vagina of vulvovaginal candidiasis（VVC） , and to draw the growth curve of lactobacillus casei, lactobacillus gasseri, lactobacillus jensenii, lactobacillus fermentum isolated from the vagi- nas of vulvovaginal candidiasis and to detection acid production and H2O2 production of them;and then to analysis the functional changes of vaginal lactobacillus in VVC and to provide a theoretical basis for the pathogeny and treatment of VVC. Methods Lactobacillus isola- ted from the vaginas of vulvovaginal candidiasis were separated, purificated, and identified by molecular biology technology. Growth curve was drawn to evaluate the growth of four lactobacillus isolated from the vaginas of vulvovaginal candidiasis. The acid producing capacity was evaluated by testing pH of four Lactobacillus bacteria liquids for 24hours. Standard curve of H2O2 concentration was drew, and then the four bacteria liquid producing amount of H2O2 per half hour was quantitatively detected. Results A total of 34 strains belonging to 4 species lactobacillus were isolated from patients with VVC. There was no significant difference between patients with VVC and healthy women in the growth rate of L. cripatus, L. gasseri L. jensenii and L. fermentum （P 〉 0.05 ）. The acid producing capacity of Lactobacillus gasseri and lactobacillus plantarum isolated from healthy women was stronger than that of VVC patients （ P 〈 0.05 ）. There was no significant difference between the two groups in lactobacillus jensenii（ P 〉 0.05 ）. But the acid producing capacity of lactobacillus fermentum isolated from VVC patients was stronger than that of than healthy women（P 〈 0.05 ）. TheH2O2 producing capacity of VVC pations was significantly less than that of the healthy women（P 〈 0. 05）. Conclusion The growth rate of lactobacillus in vaginal is stable although in the vaginal of women with VVC. The acid producting function of vaginal lactobacillus is not the key for inhibiting VVC. The H2O2 producting function of vaginal lactobacillus is a key for inhibiting VVC.